Author: Rogers, J.; Schoepp, R.J.; Schröder, O.; Clements, T.L.; Holland, T.F.; Li, J.Q.; Li, J.; Lewis, L.M.; Dirmeier, R.P.; Frey, G.J.; Tan, X.; Wong, K.; Woodnutt, G.; Keller, M.; Reed, D.S.; Kimmel, B.E.; Tozer, E.C.
Title: Rapid discovery and optimization of therapeutic antibodies against emerging infectious diseases Document date: 2008_5_13
ID: xkx56h0o_29
Snippet: A 5 0 -biotinylated dsDNA hook fragment was attached to streptavidin-coated magnetic beads. Full-length variable domains were synthesized by ligating the pool of framework 1 to the hook, washing unbound material, ligating CDR 1, followed by the pool of framework 2, and so on until all fragments were added in the correct order. Full-length reassembly products were separated from the beads with the appropriate restriction enzymes and cloned into pC.....
Document: A 5 0 -biotinylated dsDNA hook fragment was attached to streptavidin-coated magnetic beads. Full-length variable domains were synthesized by ligating the pool of framework 1 to the hook, washing unbound material, ligating CDR 1, followed by the pool of framework 2, and so on until all fragments were added in the correct order. Full-length reassembly products were separated from the beads with the appropriate restriction enzymes and cloned into pCEP4 vectors containing a mammalian secretion signal and either the kappa light chain constant domain or the IgG1 constant domain. Ligation products were transformed into E. coli (XL1Blue) and sequenced.
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